Cell Protective Activity Research Articles

Changes in cell ultra-structure, membrane permeability and protective enzyme activity in <I>Eriobotrya japonica</I> Lindl. leaves under cold stress

Changes in cell ultra-structure, membrane permeability and protective enzyme activity in <I>Eriobotrya japonica</I> Lindl. leaves under cold stress

Application of Metabolomics in the Analysis of Manufacturing Type of Pu-erh Tea and Composition Changes with Different Postfermentation Year

Liquid chromatography-mass and multivariate analyses were employed to measure the composition of pu-erh teas and to determine the general changes in the compositional patterns of pu-erh teas during postfermentation. Principle component analysis of pu-erh teas indicated two large distinct clusters in the score plot: ripened pu-erh teas and raw pu-erh teas. The raw pu-erh teas contained more antioxidant compounds compared to ripened pu-erh teas. As a result, the raw pu-erh teas showed significantly higher antioxidant activities than the ripened pu-erh teas in the 1,1-diphenyl-2-picrylhydrazyl, Trolox equivalent antioxidant capacity, and ferric reducing antioxidant power assays. In addition, raw pu-erh teas showed significantly higher NO inhibitory and cell protective activities than the ripened pu-erh teas. Significant correlations between compounds and postfermentation year were observed in raw pu-erh teas; epigallocatechin-3-gallate, epigallocatechin, epicatechin-3-gallate, and quinic acid were decreased and gallic acid was increased in a year-dependent manner. The antioxidant activity was shown to decrease as the number of antioxidant compounds in raw pu-erh tea decreased. These findings indicate that a metabolomic approach is a useful tool for analyzing manufacturing type, postfermentation year, and antioxidant activity of pu-erh tea.

Functional analyses indicate a pathogenic role of factor H autoantibodies in atypical haemolytic uraemic syndrome

Atypical haemolytic uraemic syndrome (aHUS) is associated with defective complement regulation. Recently, an autoimmune aHUS form has been described that is associated with complement factor H (CFH) autoantibodies. The aim of this study was to address the pathologic relevance of CFH autoantibodies in aHUS. CFH autoantibodies were identified and antibody levels were analysed in three aHUS patients during the disease course by the ELISA method. Epitope mapping was performed using recombinant factor H fragments and domain-mapped monoclonal antibodies. The effect of the antibodies on cell-protective activity of CFH was measured by haemolytic assays. CFH:autoantibody complexes were analysed by ELISA. All three autoantibodies bound to the C-terminal domain of CFH, which is essential for CFH binding to cell surfaces. In patient 1, plasma exchanges and immune adsorption temporarily reduced the autoantibody titre and led to temporary clinical improvement. In patient 2, plasma exchanges and long-term immunosuppression strongly reduced the CFH autoantibody level, and induced a stable remission of aHUS. Patient 3 had lower autoantibody levels that decreased during the follow-up and is in good clinical condition. The patients' plasma samples caused enhanced lysis of sheep erythrocytes, and the degree of lysis correlated with the CFH autoantibody titre and the amount of CFH:autoantibody complexes. An addition of purified CFH to aHUS plasma or removal of IgG inhibited the haemolytic activity. These results support a direct role of the autoantibodies in aHUS pathology by inhibiting the regulatory function of CFH at cell surfaces and suggest that reduction of the autoantibody titre is beneficial for the patients.

Hypoglycemic and beta cell protective effects of andrographolide analogue for diabetes treatment

BackgroundWhile all anti-diabetic agents can decrease blood glucose level directly or indirectly, few are able to protect and preserve both pancreatic beta cell mass and their insulin-secreting functions. Thus, there is an urgent need to find an agent or combination of agents that can lower blood glucose and preserve pancreatic beta cells at the same time. Herein, we report a dual-functional andrographolide-lipoic acid conjugate (AL-1). The anti-diabetic and beta cell protective activities of this novel andrographolide-lipoic acid conjugate were investigated.MethodsIn alloxan-treated mice (a model of type 1 diabetes), drugs were administered orally once daily for 6 days post-alloxan treatment. Fasting blood glucose and serum insulin were determined. Pathologic and immunohistochemical analysis of pancreatic islets were performed. Translocation of glucose transporter subtype 4 in soleus muscle was detected by western blot. In RIN-m cells in vitro, the effect of AL-1 on H2O2-induced damage and reactive oxidative species production stimulated by high glucose and glibenclamide were measured. Inhibition of nuclear factor kappa B (NF-κB) activation induced by IL-1β and IFN-γ was investigated.ResultsIn alloxan-induced diabetic mouse model, AL-1 lowered blood glucose, increased insulin and prevented loss of beta cells and their dysfunction, stimulated glucose transport protein subtype 4 (GLUT4) membrane translocation in soleus muscles. Pretreatment of RIN-m cells with AL-1 prevented H2O2-induced cellular damage, quenched glucose and glibenclamide-stimulated reactive oxidative species production, and inhibited cytokine-stimulated NF-κB activation.ConclusionWe have demonstrated that AL-1 had both hypoglycemic and beta cell protective effects which translated into antioxidant and NF-κB inhibitory activity. AL-1 is a potential new anti-diabetic agent.

Monoterpene Glucosides from Paeonia lactiflora

Four new "cage-like" monoterpene glucosides (1-4) were isolated from Paeonia lactiflora. The structures of these compounds were established by spectroscopic methods, mainly 1D and 2D NMR, and mass spectrometric analysis. Compound 4 exhibited moderate cell-protective activity against hydrogen peroxide-induced PC12 cell damage.

Phenolic compounds with cell protective activity from the fruits of Livistona chinensis

Two new depsidones, livistones A (1) and B (2), and a new benzofurane, livistone C (3), together with the 11 known compounds including three stilbenes (4–6), four steroids, three flavan-3-ols, and an alkaloid were isolated from the fruits of Livistona chinensis. The structures of the new compounds were determined by spectroscopic methods. Compounds 1, 4–6 exhibited remarkable cell protective activities against H2O2-induced SH-SY5Y cell damage.

Syntheses of tricyclic pyrones and pyridinones and protection of Aβ-peptide induced MC65 neuronal cell death

The SβC gene is conditionally expressed a 99-residue carboxy terminal fragment, C99, of amyloid precursor protein in MC65 cells and causes cell death. Consequently, MC65 cell line was used to identify inhibitors of toxicity related to intracellular amyloid β (Aβ) oligomers. Compounds that reduce the level of Aβ peptides, prevent Aβ aggregation, or eliminate existing Aβ aggregates may be used in the treatment of Alzheimer’s disease (AD). Previously, we found that a tricyclic pyrone (TP) molecule, compound 1, prevents MC65 cell death and inhibits Aβ aggregation. Hence various TPs containing heterocycle at C7 side chain and a nitrogen at position 2 or 5 were synthesized and their MC65 cell protective activities evaluated. TPs containing N3′-adenine moiety such as compounds 1 and 11 are most active with EC 50 values of 0.31 and 0.35 μM, respectively. EC 50 values of tricyclic N5-analog, pyranoisoquinolinone 13, and N2-analog, pyranopyridinone 20, are 2.49 and 1.25 μM, respectively, despite the lack of adenine moiety. Further investigation of tricyclic N2- and N5-analogs is warranted.

Functional studies indicate a direct pathogenic role of autoantibodies in atypical haemolytic uraemic syndrome by inhibiting factor H cell protective activity

Functional studies indicate a direct pathogenic role of autoantibodies in atypical haemolytic uraemic syndrome by inhibiting factor H cell protective activity

Novel Terphenyls and 3,5-Diaryl Isoxazole Derivatives Endowed with Growth Supporting and Antiapoptotic Properties

A new study on terphenyl and diaryl-isoxazole and -isoxazoline derivatives, maintaining a common 3-adamantyl-4-hydroxyphenyl moiety, has been conducted to find compounds with growth supporting and antiapoptotic properties. Unexpectedly, diphenyisoxazole derivatives bearing a nitro group replacing the carboxylic function have been found with the highest cell protective activity within the series, in complete and in serum-free conditions. Inhibition of apoptosis induced by daunorubicin has also been observed for the most active compound.

Carnosine interaction with nitric oxide and astroglial cell protection

The neuropeptide carnosine (beta-amyloid peptide aggregation has been demonstrated. Carnosine protection against peroxynitrite damage is particularly relevant, but until now there has been no evidence of any direct interaction with nitric oxide. In this study we examined the protection that carnosine provides against nitric oxide (NO)-induced cell death in primary rat astroglial cell cultures treated with lipopolysaccharide (LPS) and interferon gamma (INFgamma), a well-known neurotoxic proinflammatory condition. A correlation was found between cell protection and NO free-radical scavenging activity of carnosine. Moreover, by competitive spectrophotometric measurement and electrospray mass spectrometry analysis in cell-free experiments, we demonstrated a direct interaction of the dipeptide with NO. A comparison of carnosine with its homologues or derivatives (homocarnosine and carcinine) as well as with its amino acid constituents (L-histidine and beta-alanine) highlighted that only histidine showed significant scavenging activity. Therefore, carnosine shows direct NO-trapping ability and may be a valuable multifunctional molecule in the treatment of neurodegenerative disorders.

Pharmacologic Profile and Therapeutic Potential of NCX 4016, a Nitric Oxide‐releasing Aspirin, for Cardiovascular Disorders

NCX 4016, 2-(acetyloxy)benzoic acid 3-[(nitrooxy)methyl]phenyl ester, is a new molecule in which a nitric oxide (NO)-releasing moiety is covalently linked to aspirin. After enzymatic metabolism, NCX 4016 releases both components. In vitro and in some animal models, these components exert their pharmacologic effects simultaneously. Nitric oxide (NO) is a small gaseous molecule that exerts several activities which may prevent atherothrombotic disorders. Moreover, it displays a protective activity on the gastric mucosa. NCX 4016 has been shown to inhibit platelet activation in vitro more effectively than aspirin, to inhibit smooth muscle cell proliferation, to exert an endothelial cell protective activity and to suppress the function of several inflammatory cells potentially involved in atherothrombosis. In animal models, NCX 4016 protected from platelet thromboembolism, prevented restenosis in atherosclerosis-prone animals, protected the heart from ischemia/reperfusion injury, and induced neoangiogenesis in critically ischemic limbs. Moreover, it displayed little or no gastric toxicity and appeared to protect stomach from noxious stimuli, including aspirin. NCX 4016 has been evaluated in healthy volunteers and found to inhibit platelet cyclo-oxygenase-1 (COX-1) similarly to or slightly less than aspirin, to raise the circulating levels of NO-degradation products, and to have little or no gastric toxicity in short term studies. In particular, in phase II studies, NCX 4016 had favorable effects on effort-induced endothelial dysfunction in intermittent claudication and on platelet-activation parameters elicited by short-term hyperglycemia in type II diabetics. In patients with type II diabetes the effects of NCX 4016 on microalbuminuria and on some hemodynamic parameters were promising. The pharmacokinetics of in vivo aspirin- and NO- released by NCX 4016, as well as the bioavailability of the two molecules, were not yet adequately studied. Also, the long-term tolerability of NCX 4016, as well as its possible effectiveness in preventing ischemic cardiovascular events and progression of atherosclerosis, should be explored.

Phenolic phytochemicals in cabbage inhibit amyloid β protein-induced neurotoxicity

The antioxidative and neuronal protective effects of red and white cabbages were investigated against amyloid β protein (A β)-induced cytotoxicity in neuron-like PC12 cells. Red cabbage phenolic phytochemicals showed higher antioxidative effect. In cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), red cabbage showed more protective effect on A β-induced neurotoxicity. Since oxidative stress causes membrane breakdown, the protective effect of cabbage phytochemicals was investigated by lactate dehydrogenase and trypan blue exclusion assays. Red cabbages showed more efficient cell protective effects among samples. The overall relative neuronal cell protection activity of red cabbages by three tests was much higher than white cabbages. The degree of neuronal cell protection was closely correlated with their phenolic contents and their antioxidative capacities. Moreover, anthocyanins in cabbage appeared to be the major contributors.

Protective Effects of Daidzein on Oxidative Stress-Induced Neurotoxicity and Scopolamine-Mediated Cognitive Defect

ABSTRACT: To investigate the effect of daidzein, a major isoflavone constituent isolated from Pueraria thunbergiana (an arrowroot), on the oxidative damage and the scopolamine-induced cognitive deficit, in vitro cell viability assays and in vivo behavioral tests were performed. Using 3 assays 3-(4,5-dimethylthiazol-2-yl)–2,5-diphenyl tetrazolium bromide (MTT) reduction, lactate dehydrogenase release, and trypan blue exclusion) daidzein showed cell protective effects on oxidative stress-induced cytotoxicity. Administration of daidzein to mice significantly reversed the scopolamine-induced learning and memory impairment in a passive avoidance test. Injection of scopolamine to mice impaired performance in the passive avoidance test (43% decrease). In contrast, mice treated with daidzein before scopolamine injection were protected from this change (higher than the control value). These results suggest that daidzein's neuronal cell protective activities reduce the scopolamine-induced cognitive deficit and that its anti-amnesic effect might be due to the activation of choline acetyltransferase.

Strawberry and Its Anthocyanins Reduce Oxidative Stress-Induced Apoptosis in PC12 Cells

Because strawberries are known to contain higher concentrations of phytochemicals and have higher antioxidant capacity among common fruits, their neuroprotective activity was tested in vitro on PC12 cells treated with H2O2. Their protective effect and antioxidant capacity were also compared with those of banana and orange, which are the fresh fruits consumed at highest levels in the United States. The cell viability test using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay showed that strawberry phenolics significantly reduced oxidative stress-induced neurotoxicity. Because oxidative stress is also known to increase neuronal cell membrane breakdown, lactate dehydrogenase (LDH) and trypan blue exclusion assays were also performed. Strawberry showed the highest cell protective effects among the samples. The overall relative neuronal cell protective activity of three fruits by three tests followed the decreasing order strawberry > banana > orange. The protective effects appeared to be due to the higher phenolic contents including anthocyanins, and anthocyanins in strawberries seemed to be the major contributors.

Orally administered recombinant human interleukin-11 is protective in experimental neutropenic sepsis.

Recombinant human interleukin (IL)-11 is a multifunctional cytokine with hematopoietic, immunomodulatory, and epithelial cell protective activities. IL-11alpha receptors are expressed on the luminal surface of intestinal epithelial cells. It was hypothesized that orally administered IL-11 would prevent mucosal damage and protect against microbial invasion in a neutropenic rat model of gram-negative sepsis. IL-11 was administered daily by enteric, coated multiparticle pellets over the course of chemotherapy-induced neutropenia. Compared with the placebo group, IL-11-treated rats retained mucosal mass and had prolonged survival time, reduced pathologic changes, and reduced systemic levels of bacterial endotoxin and concentrations of Pseudomonas aeruginosa in target tissues. Enterocyte messenger RNA levels for tumor necrosis factor-alpha and interferon-gamma revealed that oral IL-11 reduced but did not prevent increased expression of these cytokine genes. These results indicate that orally administered IL-11 may preserve epithelial cell integrity in the presence of cytoreductive chemotherapy. This may represent a new treatment strategy for the prevention of infection in neutropenic hosts.

A monomeric human C4b-binding protein (C4bp) more efficiently inactivates C3b than natural C4bp: Participation of C-terminal domains in factor I-cofactor activity

We designed a cDNA construct encoding an artificial membrane molecule consisting of all 8 short consensus repeats (SCRs) of human monomeric C4b-binding protein (C4bp) followed by DAFs GPI anchor, named mC4bp, and expressed the protein on swine endothelial cells (SEC). At the same level of expression, mC4bp protected host cells as effectively as DAF, the most potent complement (C) regulator on the membrane. This result was unexpected from the reported functional properties of natural multimeric C4bp. Here, we investigated the mechanism whereby mC4bp has potent cell-protective activity. Our results were as follows: (1) mC4bp serves more efficiently as a methylamine-treated C3 (C3ma)-inactivating factor I-cofactor than natural C4bp and as efficiently as MCP as a methylamine-treated (C4ma)-inactivating cofactor by fluid-phase cofactor assay; (2) the potency of C3ma inactivation by mC4bp and factor I is quite high compared to those of other cofactors; (3)blocking studies using mAbs against C4bp suggested that both the 48 kDa N-terminal fragment and the C-terminal domain near the portion responsible for bundle formation participate in the high C3ma-inactivating capacity of mC4bp. Thus, acquiring high C3ma-inactivating capacity secondary to monomeric alteration leads to high C regulatory activity of mC4bp. These results infer that mC4bp differs from C4bp in its potent factor I-cofactor activity and is a good candidate as a safeguard against hyperacute rejection of xenografts.

Diversity of Sites for Measles Virus Binding and for Inactivation of Complement C3b and C4b on Membrane Cofactor Protein CD46

The complement system membrane cofactor protein (MCP) CD46 serves as a C3b/C4b inactivating factor for the protection of host cells from autologous complement attack and as a receptor for measles virus (MV). MCP consists of four short consensus repeats (SCR) which are the predominant extracellular structural motif. In the present study, we determined which of the four SCR of MCP contribute to its function using Chinese hamster ovary cell clones expressing each SCR deletion mutants. The results were as follows: 1) SCR1 and SCR2 are mainly involved in MV binding and infection; 2) SCR2, SCR3, and SCR4 contribute to protect Chinese hamster ovary cells from human alternative complement pathway-mediated cytolysis; and 3) SCR2 and SCR3 are essential for protection of host cells from the classical complement pathway. These results on cell protective activity of the mutants against the human classical and the alternative complement pathways were compatible with factor I-mediated inactivation profiles of C4b and C3b, respectively, in the fluid-phase assay using solubilized mutants and factor I; the results were mostly consistent with those reported by Adams et al. (Adams, E. M., Brown, M. C., Nunge, M., Krych, M., and Atkinson, J. P. (1991) J. Immunol. 147, 3005-3011). SCR2 and SCR3 were required for C3b and C4b inactivation, and SCR4-deleted MCP showed weak cofactor activity for C4b cleavage but virtually no cofactor activity for C3b cleavage. The functional domains of MCP for the three natural ligands C3b, C4b, and MV, therefore, map to different, although partly overlapping, SCR domains.

Membrane cofactor protein (CD46) protects cells predominantly from alternative complement pathway-mediated C3-fragment deposition and cytolysis.

Membrane cofactor protein (MCP) cDNA was transfected into Chinese hamster ovarian tumor (CHO) cells and the functional properties of the expressed protein were studied. Cells adherent to flasks were essential for continuous expression of MCP on CHO cells. If the cells were maintained in noncoated flasks, MCP expression was markedly reduced but upon being transferred to coated flasks reexpressed the protein. MCP expressed on CHO cells had the expected m.w., approximately 50 kDa, and possessed factor I-cofactor activity. By a propidium iodide incorporation assay and by 51Cr release assay, antibody-sensitized CHO cells expressing MCP were protected from C-mediated cytotoxicity. The inhibition of lytic activity correlated with a decrease in C3 deposition. This host cell protective activity was exerted efficiently for the alternative pathway. The classical pathway was not blocked by MCP unless the cells were presensitized with low concentrations of antibody. These results imply that MCP primarily protects host cells from alternative pathway-mediated C3 targeting. In a pathologic state such as autoimmune diseases, the binding of an autoantibody to a target may overcome this protective effect of MCP via the classical pathway.

Cell-protective effect of human saliva specific for herpes simplex virus

Saliva from normal human adults pre-incubated with human amnion cell cultures protected these cells against destruction by herpes simplex virus. In two-thirds of subjects tested, protection amounted to plaque reduction of 55–94 per cent. Protection seemed to be achieved by direct action on cells rather than inactivation of virus. Evidence indicates that the active material was not antibody, although it was more active against herpes simplex virus than vaccinia or vesicular stomatitis virus. This cell-protective activity correlated with recurrent herpes labialis and may be significant in the varying resistance of individuals to symptomatic recurrences and may explain the tendency of recurrent lesions to involve the lips and circumoral skin although sparing the oral mucous membrane.